Abstract
Immobilised metal-ion affinity chromatography (IMAC) is widely used for the purification of recombinant proteins in which a poly-histidine tag is introduced. However, other proteins may also bind to IMAC columns. We describe the use of a washing buffer with a low concentration of EDTA (0.5 mM) for the removal of proteins without histidine tag from IMAC columns. Four histidine-tagged recombinant proteins/protein complexes were purified to homogeneity from cell culture medium of insect cells by using an EDTA washing buffer. The presence of a low concentration of EDTA in washing buffers during IMAC may have a general application in the purification of histidine-tagged proteins. (C) 2001 Elsevier Science B.V. All rights reserved.
| Original language | English |
|---|---|
| Pages (from-to) | 129-136 |
| Number of pages | 8 |
| Journal | Journal of Chromatography B |
| Volume | 760 |
| Issue number | 1 |
| Publication status | Published - 25-Aug-2001 |
Keywords
- recombinant proteins
- EDTA
- histidine
- CONTAINING CYTOCHROME-C
- VIRUS
- GLYCOPROTEIN
- GH
- COMPLEX
- BINDING
- GL